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A simple and efficient CRISPR/Cas9 system permits ultra-multiplex genome editing in plants

文献类型： 外文期刊

作者： Wu, Suting 1 ; Kyaw, Htin 1 ; Tong, Zhijun 6 ; Yang, Yirong 1 ; Wang, Zhiwei 1 ; Zhang, Liying 1 ; Deng, Lihua 4 ; Zhang, Zhiguo 1 ; Xiao, Bingguang 6 ; Quick, William Paul 1 ; Lu, Tiegang 1 ; Xiao, Guoying 4 ; Qin, Guannan 2 ; Cui, Xue'an 1 ;

作者机构： 1.Chinese Acad Agr Sci, Biotechnol Res Inst, Beijing 100081, Peoples R China

2.Fujian Agr & Forestry Univ, Coll Agr, Key Lab Genet Breeding & Multiple Utilizat Crops, Minist Educ, Fuzhou 350002, Fujian, Peoples R China

3.Fujian Agr & Forestry Univ, Coll Agr, Minist Agr & Rural Affairs, Key Lab Biol Breeding Fujian & Taiwan Crops, Fuzhou 350002, Fujian, Peoples R China

4.Chinese Acad Sci, Key Lab Agroecol Proc Subtrop Reg, Inst Subtrop Agr, Changsha 410125, Hunan, Peoples R China

5.Univ Sheffield, Sch Biosci, Firth Court, Sheffield S10 2TN, England

6.Yunnan Acad Tobacco Agr Sci, Natl Tobacco Genet Engn Res Ctr, Key Lab Tobacco Biotechnol Breeding, Kunming 650021, Yunnan, Peoples R China

关键词： CRISPR/Cas9; Multiplex genome editing; Assembly system; Plant

期刊名称：CROP JOURNAL （ 影响因子：6.6； 五年影响因子：6.5 ）

ISSN： 2095-5421

年卷期： 2024 年 12 卷 2 期

页码：

收录情况： SCI

摘要： The development and maturation of the CRISPR/Cas genome editing system provides a valuable tool for plant functional genomics and genetic improvement. Currently available genome-editing tools have a limited number of targets, restricting their application in genetic research. In this study, we developed a novel CRISPR/Cas9 plant ultra-multiplex genome editing system consisting of two template vectors, eight donor vectors, four destination vectors, and one primer-design software package. By combining the advantages of Golden Gate cloning to assemble multiple repetitive fragments and Gateway recombination to assemble large fragments and by changing the structure of the amplicons used to assemble sgRNA expression cassettes, the plant ultra-multiplex genome editing system can assemble a single binary vector targeting more than 40 genomic loci. A rice knockout vector containing 49 sgRNA expression cassettes was assembled and a high co-editing efficiency was observed. This plant ultra-multiplex genome editing system advances synthetic biology and plant genetic engineering. (c) 2024 Crop Science Society of China and Institute of Crop Science, CAAS. Production and hosting by Elsevier B.V. on behalf of KeAi Communications Co., Ltd. This is an open access article under the CC BY-NCND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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